摘要
目的探讨金欣康颗粒(丹参、桂枝、黄芪、人参、毛冬青等7味药组成)对心力衰竭(简称心衰)大鼠线粒体自噬蛋白LC3/Beclin1表达的影响,阐述金欣康颗粒改善心梗后心衰心肌纤维化的机制。方法复制心梗后心衰大鼠模型。将造模成功的心衰大鼠随机分为4组:模型组,金欣康颗粒低剂量组、高剂量组和培哚普利组,另设假手术组,每组10只。给药12周,以HE染色、天狼猩红染色评估心肌组织的病理变化,心脏彩超对心脏结构和心功能进行评估,透射电镜观察自噬小体,免疫组化法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型和Ⅲ型胶原(CollagenⅠ和CollagenⅢ)蛋白的表达,Western Blot法检测自噬蛋白LC3、Beclin1的表达。结果成功复制心梗后心衰大鼠模型,模型组大鼠梗死区周边区域有大量红色的胶原沉积,心肌纤维排列紊乱,部分纤维断裂;线粒体明显肿胀,出现峭断裂,能观察到较多的自噬小体。金欣康颗粒组和培哚普利组心肌纤维排列较模型组整齐,纤维化程度较轻,自噬小体更加明显。与假手术组比较,模型组的射血分数(LVEF)和短轴缩短率(LVFS)显著降低(P<0.01),左室舒张末期内径(LVEDd)及收缩末期内径(LVEDs)增大(P<0.01),α-SMA、CollagenⅠ和CollagenⅢ表达水平升高(P<0.05),线粒体自噬蛋白Beclin1和LC3Ⅱ/LC3Ⅰ的表达升高(P<0.05);与模型组比较,金欣康颗粒组和培哚普利组LVEF和LVFS明显提高(P<0.01),心功能得到改善;α-SMA、CollagenⅠ和CollagenⅢ蛋白表达降低(P<0.05),Beclin1和LC3Ⅱ/LC3Ⅰ的表达升高(P<0.05)。结论金欣康颗粒可减轻心梗后心衰大鼠的心肌纤维化程度,其机制可能与增强线粒体自噬蛋白LC3Ⅱ/LC3Ⅰ和Beclin1的表达有关。
Objective To explore the effect of Jinxinkang Granules(composed of Salviae Miltiorrhizae Radix et Rhizoma,Cinnamomi Ramulus,Astragali Radix,Ginseng Radix et Rhizoma,Ilies Pubescentis Radix,etc.)on the expression of mitochondrial autophagy protein LC3/Beclin1,and to clarify the mechanism of Jinxinkang Granules on improving myocardial fibrosis after myocardial infarction.Methods A SD rat model of postinfarction heart failure was constructed.Successful model rats were randomly divided into 4 groups:the model group(Model),Perindopril group(PDPL),Jinxinkang Granules low-dose(JXK-L)and high-dose(JXK-H)groups.A sham operation group(Sham)was also established,each group had 10 rats.After 12-week administration of drug,pathological changes of myocardial tissue were detected by using HE and Sirius red staining.Cardiac structure and function were assessed by echocardiography.Transmission electron microscopy was used to observe autophagic bodies,and immunohistochemistry was adopted to detect the expressions ofα-smooth muscle actin(α-SMA),collagen typeⅠ(CollagenⅠ)and collagen typeⅢ(CollagenⅢ).Western Blot was also performed to detect the expressions of autophagy proteins LC3 and Beclin1.Results The rat model of postinfarction heart failure was successfully constructed.In the model group,we found large amounts of red collagen deposits in peripheral area of infarction,disorganized myocardial fibers and breaks of myocardial fibers.Mitochondria appeared swollen and disrupted cristae.Quite a few autophagic vesicles could be observed.The arrangement of myocardial fibers in JXK group and PDPL group were more neatly than those in the model group.Meanwhile,the degree of myocardial fibrosis was mild,and more autophagic vesicles were observed.Compared with the Sham group,left ventricular ejection fraction(LVEF)and short axis shortening rate(LVFS)were significantly decreased in the model group(P<0.01),and left ventricular end diastolic diameter(LVEDd)and end systolic diameter(LVEDs)were increased(P<0.01),the levels ofα-SMA,CollagenⅠand CollagenⅢwere significantly increased(P<0.05),and the expressions of Beclin1 and LC3Ⅱ/LC3Ⅰwere also raised(P<0.05).Compared with the model group,LVEF and LVFS were significantly increased in the JXK and PDPL groups(P<0.01),and cardiac function was improved.The protein expressions ofα-SMA,CollagenⅠand CollagenⅢwere significantly decreased(P<0.05).The Beclin1 and LC3Ⅱ/LC3Ⅰexpressions were significantly increased(P<0.05).Conclusions Jinxinkang Granules can reduce the degree of myocardial fibrosis in rats with postinfarction heart failure,and its mechanism may be related to enhancing the expressions of mitochondrial autophagy proteins LC3Ⅱ/LC3Ⅰand Beclin1.
作者
赵新军
黎燚华
康亮
陈琦
林莉雯
褚庆民
罗川晋
彭锐
都治伊
李荣
ZHAO Xinjun;LI Yihua;KANG Liang;CHEN Qi;LIN Liwen;CHU Qingmin;LUO Chuanjin;PENG Rui;DU Zhiyi;LI Rong(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2023年第6期713-721,共9页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
广东省基础与应用基础项目(2021A1515220046)
广州市科技计划项目(202201020356)
广州中医药大学“双一流”与高水平大学学科协同创新团队项目(A1-2601-21-415-021)
作者简介
赵新军,男,博士,副主任医师,研究方向:中西医结合防治心衰心室重构。Email:zxjhwxm@sina.com;通信作者:李荣,男,博士,主任医师,研究方向:心衰病机及中医药对心衰和病态窦房结综合征的防治。Email:lrhbs@126.com。
